Isorhapontigenin Modulates SOX9/TOLLIP Expression to Attenuate Cell Apoptosis and Oxidative Stress in Paraquat-Induced Acute Kidney Injury

Figure 3

SRY-box transcription factor 9 (SOX9) regulates Toll-interacting protein (TOLLIP) expression and attenuates paraquat (PQ) toxicity to NRK-52E cells. NRK-52E cells were pretreated with ISO (5 μM or 10 μM) for 30 min followed by a 24 h coincubation with 300 μM PQ. The expression of SOX9 and TOLLIP was evaluated at (a, b) protein and (c) mRNA levels using Western blot and quantitative real-time PCR (qPCR). (d) A dual-luciferase reporter assay was performed to explore the effects of SOX9 on the activities of the TOLLIP promoter. (e, f) Verification of adenovirus- (Ad-) mediated overexpression of SOX9 in NRK-52E cells using Western blot and qPCR. Forty-eight hours after Ad transduction, cells were incubated with 300 μM PQ for 24 h. (g, h) Flow cytometry was performed to determine the cell apoptosis. Levels of (i) malondialdehyde (MDA) and (j) lactate dehydrogenase (LDH) were measured using the commercially available kits. TOLLIP expression was determined at the (k) protein and (l) mRNA levels using Western blot and qPCR. Error bars represent standard deviations. values < 0.05, values < 0.01, and values < 0.001.

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